Abstract
Fluorescent D-amino acids (FDAAs) enable in situ visualization of bacterial cell wall synthesis via their incorporation into peptidoglycan (PG) crosslinks. When combined with super-resolution microscopy, FDAAs allow the details of cell wall synthesis to be resolved beyond the diffraction limit of visible light. Here, we describe using the super-resolution method of single-molecule localization microscopy (SMLM) in conjunction with two newly synthesized FDAAs (sCy5DA and sCy5DL_amide) to resolve bacterial PG at the nanoscale in a variety of species, including Gram-negative, Gram-positive, and mycobacteria.
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Acknowledgments
This work was supported by the European Union’s H2020 program under the European Research Council (ERC; CoG 819823 Piko, to S.M. and C.Z.). We thank Matthew D. Lycas for giving suggestions on this protocol after observing experiments.
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Zhang, C., Manley, S. (2024). Super-Resolution Microscopy of the Bacterial Cell Wall Labeled by Fluorescent D-Amino Acids. In: Ton-That, H. (eds) The Bacterial Cell Wall. Methods in Molecular Biology, vol 2727. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3491-2_7
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DOI: https://doi.org/10.1007/978-1-0716-3491-2_7
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